Use of a new multiplex quantitative polymerase chain reaction based assay for simultaneous detection of Neisseria meningitidis, Escherichia coli K1, Streptococcus agalactiae, and Streptococcus pneumoniae
Background and Objectives: Neisseria meningitidis, Escherichia coli K1, Streptococcus agalactiae, and Streptococcus pneumoniae cause 90% of bacterial meningitis. Almost all infected people die or have irreversible neurological complications. Therefore, it is essential to have a diagnostic kit with the ability to quickly detect these fatal infections.
Materials and Methods: The project involved 212 patients from whom cerebrospinal fluid samples were obtained. After total genome extraction and performing multiplex quantitative polymerase chain reaction (qPCR), the presence or absence of each infectious factor was determined by comparing with standard strains.
Results: The specificity, sensitivity, positive predictive value, and negative predictive value calculated were 100%, 92.9%, 50%, and 100%, respectively. So, due to the high specificity and sensitivity of the designed primers, they can be used instead of bacterial culture that takes at least 24 to 48 hours.
Conclusion: The remarkable benefit of this method is associated with the speed (up to 3 hours) at which the procedure could be completed. It is also worth noting that this method can reduce the personnel unintentional errors which may occur in the laboratory. On the other hand, as this method simultaneously identifies four common factors that cause bacterial meningitis, it could be used as an auxiliary method diagnostic technique in laboratories particularly in cases of emergency medicine.
2. Principi N, Esposito S. Bacterial meningitis: new treatment options to reduce the risk of brain damage. Expert Opin Pharmacother 2020;21: 97-105.
3. Obaro S. Updating the diagnosis of bacterial meningitis. Lancet Infect Dis 2019;19: 1160-1161.
4. Kourna Hama M, Khan D, Laouali B, Okoi C, Yam A, Haladou M, et al. Pediatric bacterial meningitis surveillance in Niger: increased importance of Neisseria meningitidis serogroup C, and a decrease in Streptococcus pneumoniae following 13-valent pneumococcal conjugate vaccine introduction. Clin Infect Dis 2019;69(Suppl 2): S133-S139.
5. Jing-Zi P, Zheng-Xin H, Wei-Jun C, Yong-Qiang J. Detection of bacterial meningitis pathogens by PCR-Mass spectrometry in cerebrospinal fluid. Clin Lab 2018;64: 1013-1019.
6. Sacchi CT, Fukasawa LO, Gonçalves MG, Salgado MM, Shutt KA, Carvalhanas TR, et al. Incorporation of real-time PCR into routine public health surveillance of culture negative bacterial meningitis in São Paulo, Brazil. PLoS One 2011;6(6): e20675.
7. Papavasileiou K, Papavasileiou E, Tzanakaki G, Voyatzi A, Kremastinou J, Chatzipanagiotou S. Acute bacterial meningitis cases diagnosed by culture and PCR in a children’s hospital throughout a 9-Year period (2000-2008) in Athens, Greece. Mol Diagn Ther 2011;15: 109-113.
8. Nigrovic LE, Malley R, Macias CG, Kanegaye JT, Moro-Sutherland DM, Schremmer RD, et al. Effect of antibiotic pretreatment on cerebrospinal fluid profiles of children with bacterial meningitis. Pediatrics 2008;122: 726-730.
9. Bryan JP, Desilva HR, Tavares A, Rocha H, Scheld WM. Etiology and mortality of bacterial meningitis in northeastern Brazil. Rev Infect Dis 1990;12: 128-135.
10. Ragunathan L, Ramsay M, Borrow R, Guiver M, Gray S, Kaczmarski EB. Clinical features, laboratory findings and management of meningococcal meningitis in England and Wales: report of a 1997 survey. meningococcal meningitis: 1997 survey report. J Infect 2000;40: 74-79.
11. Motamedifar M, Sedigh Ebrahim-Saraie H, Mansury D, Nikokar I, Hashemizadeh Z. Prevalence of etiological agents and antimicrobial resistance patterns of bacterial meningitis in Nemazee hospital, Shiraz, Iran. Arch Clin Infect Dis 2015;10(2): e22703.
12. Fouad R, Khairy M, Fathalah W, Gad T, El-Kholy B, Yosry A. Role of clinical presentations and routine CSF analysis in the rapid diagnosis of acute bacterial meningitis in cases of negative gram stained smears. J Trop Med 2014;2014: 213762.
13. Khater WS, Elabd SH. Identification of common bacterial pathogens causing meningitis in culture-negative cerebrospinal fluid samples using real-time polymerase chain reaction. Int J Microbiol 2016;2016: 4197187.
14. Brouwer MC, Tunkel AR, van de Beek D. Epidemiology, diagnosis, and antimicrobial treatment of acute bacterial meningitis. Clin Microbiol Rev 2010;23: 467-492.
15. Eskola J. Use of conjugate vaccines to prevent meningitis caused by Haemophilus influenzae type b or Streptococcus pneumoniae. J Hosp Infect 1995;30 Suppl:313-321.
16. Brink M, Welinder-Olsson C, Hagberg L. Time window for positive cerebrospinal fluid broad-range bacterial PCR and Streptococcus pneumoniae immunochromatographic test in acute bacterial meningitis. Infect Dis (Lond) 2015;47: 869-877.
17. Wu HM, Cordeiro SM, Harcourt BH, Carvalho M, Azevedo J, Oliveira TQ, et al. Accuracy of real-time PCR, gram stain and culture for Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae meningitis diagnosis. BMC Infect Dis 2013;13: 26.
18. Afifi S, Wasfy MO, Azab MA, Youssef FG, Pimentel G, Graham TW, et al. Laboratory-based surveillance of patients with bacterial meningitis in Egypt (1998–2004). Eur J Clin Microbiol Infect Dis 2007;26: 331-340.
|Issue||Vol 13 No 4 (2021)|
|Cerebrospinal fluid; Meningitis; Quantitative polymerase chain reaction; Simultaneous detection; Diagnosis testing|
|Rights and permissions|
|This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.|